PROJECT SUMMARY/ABSTRACT This SBIR grant will drive a promising form of T cell therapy, called ATTACK, toward mainstream society. ATTACK's promise as a cancer therapy is supported by clinical data that suggest it has the capacity to effectively treat solid and liquid tumors, expose cancer cells not recognized by ATTACK to the patient's own immune system for destruction, circumvent the need for patient immunosuppression, avoid deadly toxicities associated with genetically engineering approaches such as CAR T, prime the patient's immune system to facilitate ongoing surveillance for cancer cells, and eliminate complex genetic engineering with live virus. Unfortunately, despite its unique advantages and impressive clinical results, ATTACK is currently saddled with a very complex manufacturing process that prevents commercialization. This SBIR program is intended to render the ATTACK manufacturing process commercially viable. The investigation is a two-step process. First we will create novel methods of activating cancer killing T cells to recognize multiple tumor associated antigens. Second, we will use these methods to create a commercially viable ATTACK manufacturing process. If successful, the stage will be set for Phase III commercialization of a highly advantageous cancer therapy. Aim 1 Objective: Create novel test devices for use in Aim 2 by constructing centrifuge compatible cell culture devices integrating G-Rex design principals as well as the required geometry to facilitate the activation, stimulation, and medium exchange investigation of Aim 2. ? Aim 1 Deliverable: WW will deliver fully functional, sterile, centrifuge compatible cell culture devices to BCM personnel for the activation, stimulation, and medium exchange optimization studies outlined in Aim 2. Aim 2 Objective: Identify a simple and robust manufacturing process for the generation of ATTACK T cells by undertaking a three step investigation: (i) determine if multiple antigen stimulation can be avoided by increasing the strength of the antigen stimulation signal at the onset of the culture, (ii) identify an alternative source of APCs that can replace the use of DCs, (iii) combine the knowledge obtained in (i) and (ii) to define the optimal final manufacturing protocol. ? Aim 2 Deliverable: Protocol(s) will be available for establishing the design objectives of Aim 3 and to establish the full scale production process of Aim 4. Aim 3 Objective: Based upon the knowledge gained in Aim 2, create G-Rex devices of optimal size for use in the full scale production process study of Aim 4. ? Aim 3 Deliverable: Full scale G-Rex devices will be available for use in Aim 4. Aim 4 Objective: Verify protocols optimized in Aim 2 are repeatable in full scale G-Rex production. ? Aim 4 Deliverable: Specifications for an optimal and commercially viable ATTACK manufacturing process will be defined for the Phase III commercialization effort.